USPTO Patents Application 09619290

Attorney Docket No. 0152.00378 -122-

CLAIMS

What is claimed is:

5 1. An isolated and purified glycoprotein and functional

analogues thereof characterized by:

(a) being expressed on at least primitive hematopoietic cells,

(b) being a ligand for L-selectin or E-selectin t the binding of
ligand to L-selectin or E-selecting being uninhibited by anti-CD34 antibodies;

10 (c) being resistant to O-sialoglycoprotein endopeptidase

activity;

(d) being unrecognized by MECA-79 a monoclonal antibody
which identifies ligands of L-selectin on lymph node high endothelial venules;
and

15 (e) being sulfation-independent.

2. An isolated and purified glycoprotein and functional
analogues thereof as set forth in claim 1 wherein said glycoprotein is a
membrane-associated glycoprotein.

20

3. An isolated and purified glycoprotein and functional
analogues thereof as set forth in claim 1 wherein said glycoprotein functions
as an adhesion protein ligand.

25

4. An isolated and purified glycoprotein and functional
analogues thereof as set forth in claim 1 wherein said glycoprotein facilitates
attachment of lymphocytes to hematopoietic cells.

Attorney Docket No. 0152.00378

-123-

5. At least one antibody directed against said glycoprotein and
functional analogues thereof as set forth in claim 1.

6. A method of targeting cells expressing the glycoprotein as
set forth in claim 1 including the steps of:

preparing a monoclonal antibody directed against the
glycoprotein as set forth in claim 1,

preparing an immunotoxin utilizing the antibody,

exposing a population of cells to said antibodies, and

killing cells bound to the immunotoxin.

7. The method of claim 6 wherein the toxin is selected from the
group consisting of ricin A chain, pseudomonas exotoxin A, diphtheria toxin
and chemotherapeutic compounds.

8. The method of claim 6 wherein the cells are exposed to the
immunotoxin in vivo.

9. The method of claim 6 further characterized by the cells
being selected from the group consisting of leukemic cells, malignant
hemopoietic progenitor cells and other malignant cells expressing the
glycoprotein.

10. A method of selecting for cells expressing the glycoprotein
as set forth in claim 1 including the steps of

Attorney Docket No. 01 52.00378 -1 24-

preparing an antibody directed against the glycoprotein as set
forth in claim 1,

exposing a population of cells to said antibodies, and
selecting cells bound to the antibody.

5

1 1 . The method of claim 10 wherein said selecting step is
selected from complement-mediated lysis, panning, cell sorting.

12. A method of regulating hematopoiesis including the step of:

10 selecting cells with an appropriate level of expression of the

glycoprotein as set forth in claim 1 from a patient.

culturing the selected cells, and

reinfusing the patient with the expanded selected cell

population.

15

13. A method of regulating inflammatory response by
interrupting cellular migration into lymph nodes and sites of chronic
inflammation including the step of administering to a patient functional
analogues or antibody directed against the glycoprotein as set forth in claim

20 1.

14. The method of claim 13 further characterized by the
inflammatory response being as found in the group selected from
autoimmune disorders, post-ischemic tissue injury and sepsis.

25

Attorney Docket No. 0152.00378 -125-

15. A method of performing an overlay adherence assay by
using cell suspensions as a substrate.

16. The method of claim 15 further characterized by preparing
5 the single cell suspension substrate by depositing the single cell suspension

on a slide using a modified sample chamber for use in a cytocentrifuge.

17. A method of making a cytocentrifuge sample chamber
assembly by connecting together a slide and a sample chamber, the sample

10 chamber including a cell substrate depositing port and fixing the cell substrate
depositing port at one of a plurality of positions relative to the slide.

18. The method of claim 17 whereby said step of fixing the cell
substrate depositing port at one of a plurality of positions relative to the slide

1 5 is further defined as removing a lateral edge region from an end flange of the
sample chamber thereby displacing the sample chamber laterally and/or
vertically.

19. A cytocentrifuge sample chamber assembly comprising:

20 a sample chamber including cell substrate receiving means for

receiving a cell substrate and depositing means for depositing a cell substrate
on a slide surface during cytocentrifugation; and

connecting means for connecting together a slide and said
sample chamber and fixing said depositing means at one of a plurality of
25 - positions relative to said connecting means.

Attorney Docket No. 0152.00378 -126-

20. A cytocentrifuge sample chamber assembly as set forth in
claim 19 wherein said connecting means include an end flange with unequal
sized side areas to allow lateral displacement of said depositing means.

5 21 . A cytocentrifuge sample chamber assembly as set forth in

claim 19 wherein said connecting means include a generally rectangular plate
disposed normal to and along an end flange with a removed coextensive
lower edge region of both said plate and said end flange to allow vertical
displacement of said depositing means and said connecting means include a
10 flat generally rectangular plate disposed normal to and along an end flange
with a removed coextensive lower edge region of both said plate and said end
flange and unequal sized side areas of said end flange to allow vertical and
lateral displacement of said depositing means.

15 22. A cytocentrifuge sample chamber assembly as set forth in

claim 22 wherein said receiving means include a funnel interconnected with
said discharge port such that under cytocentrifugation a cell substrate
disposed in said funnel enters said discharge port.

20 23. A kit consisting of a series of incrementally modified sample

chambers, said incremental modifications allowing incremental vertical and
lateral displacement of a sample chamber in a holder assembly thereby
allowing deposition of a cell substrate at one of a plurality of positions on a
slide.

24. A method of determining a pharmaceutical use by:
modifying L-selectin or E-selectin activity of a cell line; and

Attorney Docket No. 0152.00378 -127-

applying pharmaceuticals to said cell line which affect the
modified selectin activity.

25. The method according to claim 24, wherein said applying
5 step further includes modulating selectin activity with the pharmaceuticals.

26. A cell line expressing the glycoprotein as set forth in claim

1.

10 27. The cell line according to claim 26, wherein said cell line is

developed by positive selection.